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United States Patent | 6,165,718 |
Borchert ,   et al. | December 26, 2000 |
A method for in vivo production of a library in cells comprising a multitude of mutated genetic elements, wherein an error-prone polymerase is used in each ancestral cell to replicate all or a part of a genetic element independently of the host chromosomal replication machinery. The genetic element comprises i) an origin of replication from which replication is initiated, ii) optionally a genetic marker, e.g. a gene conferring resistance towards an antibiotic, iii) a gene encoding the polypeptide of interest. Also methods for the generation of a DNA sequence encoding a desired variant of a polypeptide of interest, and for the determination of such a DNA sequence are described.
Inventors: | Borchert; Torben Vedel (Jyllinge, DK); Ehrlich; Stanislas Dusko (Paris, FR) |
Assignee: | Novo Nordisk A/S Novo Alle (Bagsvaerd, DK) |
Appl. No.: | 112410 |
Filed: | July 8, 1998 |
Jan 10, 1996[DK] | 0018/96 |
Current U.S. Class: | 435/6; 435/69.1; 435/189; 435/193; 435/195; 435/252.33; 435/320.1; 435/471; 435/476; 435/488; 435/489 |
Intern'l Class: | C12Q 001/68; C12N 015/70; C12N 015/63; C12N 009/10; C12N 009/14; 252.33; 195; 189; 193 |
Field of Search: | 435/69.1,7.1,69.2,91.4,455,471,476,483,484,485,487,488,320.1,348,325,354,6,489 |
Foreign Patent Documents | |||
WO 90/07576 | Jul., 1990 | WO. |
James E. Bailey, Science, vol. 252, pp. 1668-1675, Jun. 21, 1991. |